In-vitro susceptibility and ex-vivo evaluation of macrocyclic lactone endectocides sub-lethal concentrations against Plasmodium vivax oocyst development in Anopheles arabiensis

Abstract

Background: Asymptomatic malaria transmission has become a public health concern across malaria-endemic Africa including Ethiopia. Specifically, Plasmodium vivax is more efficient at transmitting earlier in the infection and at lower densities than Plasmodium falciparum. Consequently, a greater proportion of individuals infected with P. vivax can transmit without detectable gametocytaemia. Mass treatment of livestock with macrocyclic lactones (MLs), e.g., ivermectin and doramectin, was suggested as a complementary malaria vector tool because of their insecticidal effects. However, the effects of MLs on P. vivax in Anopheles arabiensis has not yet been fully explored. Hence, comparative in-vitro susceptibility and ex-vivo studies were conducted to evaluate the effects of ivermectin, doramectin and moxidectin sub-lethal concentrations on P. vivax oocyst development in An. arabiensis.

Methods: The 7-day sub-lethal concentrations of 25% (LC₂₅) and 5% (LC₅) were determined from in-vitro susceptibility tests on female An. arabiensis in Hemotek® membrane feeding assay. Next, an ex-vivo study was conducted using P. vivax gametocytes infected patient's blood spiked with the LC₂₅ and LC₅ of the MLs. At 7-days post-feeding, each mosquito was dissected under a dissection stereo microscope, stained with 0.5% (w/v) mercurochrome solution, and examined for the presence of P. vivax oocysts. Statistical analysis was based on a generalized mixed model with binomially distributed error terms.

Results: A 7-day lethal concentration of 25% (LC₂₅, in ng/mL) of 7.1 (95% CI: [6.3;8.0]), 20.0 (95%CI:[17.8;22.5]) and 794.3 (95%CI:[716.4;1516.3]) were obtained for ivermectin, doramectin and moxidectin, respectively. Similarly, a lethal concentration of 5% (LC₅, in ng/mL) of 0.6 (95% CI: [0.5;0.7]), 1.8 (95% CI:[1.6;2.0]) and 53.7 (95% CI:[ 48.4;102.5]) were obtained respectively for ivermectin, doramectin and moxidectin. The oocyst prevalence in treatment and control groups did not differ significantly (p > 0.05) from each other. Therefore, no direct effect of ML endectocides on P. vivax infection in An. arabiensis mosquitoes was observed at the sub-lethal concentration (LC25 and LC5).

Conclusions: The effects of ivermectin and doramectin on malaria parasite is more likely via indirect effects, particularly by reducing the vectors lifespan and causing mortality before completing the parasite's sporogony cycle or reducing their vector capacity as it affects the locomotor activity of the mosquito.

Keywords: Anopheles arabiensis; Membrane feeding; Oocyst; Plasmodium vivax; Sublethal concentrations.

Fig. 1

Anopheles arabiensis mosquito mortality (in percentage) including the biological death after ingestion of cattle blood spiked with different concentrations (in ng/mL) of ivermectin, doramectin and moxidectin

Fig. 2

Kaplan–Meier survival curves showing the survival probability of Anopheles arabiensis mosquitoes as a function of time after feeding on cattle blood spiked with different concentration (ng/ml)) of the ML endectocides ivermectin (IVER) (A), doramectin (DORA) (B) and moxidectin (C)

Fig. 3

Plasmodium vivax oocyst prevalence (in percentage) versus dose administered in Anopheles arabiensis mosquitoes in the ivermectin, moxidectin, doramectin treated and control groups. Control refers to those administered with P. vivax infected blood without drug

Fig. 4

Oocyst intensity among Plasmodium vivax infected Anopheles arabiensis mosquitoes treated with doramectin, ivermectin, moxidectin and control groups